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An optimised micro-titer plate method for characterisation of endogenous rye phytase under industrial rye bread making conditions

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Abstract

A modified rapid method for the determination of phytase activity has been developed by measurements of molybdenum blue complex using micro-titer plate reader. This method requires only a small amount of extract, and it is possible to make more replicates at the same time and thus a better sampling and a higher capacity compared to measurements on spectrophotometer, which is traditionally used for those measurements. The extraction time was reduced by 36% compared to standard method and the extinction coefficient for molybdenum blue complex was determined instead of using P-standard solutions. To demonstrate the new method it was applied to the determination of the activity of phytase at a range of pH values and temperatures relevant to the rye bread making process. The activity of rye phytase was 320 nkatal g−1 grains (pH 5.5, 37 °C). The temperature optimum was 45–55 °C and the pH optimum 6.0. The study revealed that the rye phytase is very pH-sensitive and quite stable at different temperatures during the whole bread-making process. The enzyme activity of the endogenous rye phytase is optimal for a total degradation of phytic acid during rye bread making and thus the full bioavailability of phytate-bound minerals.

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Authors and Affiliations

  1. Quality and Technology, Department of Food Science, Faculty of Life Sciences, University of Copenhagen, Rolighedsvej 30, 1958, Frederiksberg C, Denmark

    Merete Møller Nielsen, Marianne Linde Damstrup & Åse Hansen

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  1. Merete Møller Nielsen
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  2. Marianne Linde Damstrup
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  3. Åse Hansen
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Correspondence to Åse Hansen.

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Nielsen, M.M., Damstrup, M.L. & Hansen, Å. An optimised micro-titer plate method for characterisation of endogenous rye phytase under industrial rye bread making conditions. Eur Food Res Technol 227, 1009–1015 (2008). https://doi.org/10.1007/s00217-007-0814-6

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  • DOI: https://doi.org/10.1007/s00217-007-0814-6

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